An enzymatic assay of chloramphenicol coupled with fluorescence reaction.
An alternative method for the assay of chloramphenicol using an enzymatic reaction coupled with a fluorescence detection system has been developed. Chloramphenicol was enzymatically acetylated by chloramphenicol acetyltransferase in the presence of acetyl-coenzyme A (acetyl-CoA) as the acetyl-donor, after which the liberated CoA-SH was derivatized with a fluorogenic reagent, 4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole. The assay was linear over the range of 2.5-40 micrograms/ml. Analytical recoveries of chloramphenicol at concentrations of 7.5 and 22.5 micrograms/ml, added to human serum, plasma, or a slightly hemolyzed serum, were in the range of 93.3 to 106.2%. The enzymatic assay was not affected by the presence of ten other antibiotics tested.
Yamato
S
S
Department of Analytical Chemistry, Niigata College of Pharmacy, Japan.
Sugihara
H
H
Shimada
K
K
D002701:Chloramphenicol / Q000097:blood*; D005456:Fluorescent Dyes; D006801:Humans; D010069:Oxadiazoles
D005456:Fluorescent Dyes; D010069:Oxadiazoles; D002701:Chloramphenicol; C046364:4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole